The role of uhpA in Edwardsiella piscicida and the inflammatory cytokine response in tilapia

Abstract

Edwardsiella piscicida (E. piscicida) is an important zoonotic pathogen that infects fish by colonizing the intestines. The intestine provides nutrition including Glucose 6-phosphate (Glu6P) and a competitive environment for the microbiota. Although the transport system regulatory protein gene uhpA has been reported in E. piscicida genomes, whether the uhpA gene is involved in the pathogenicity of E. piscicida remains largely unknown. Therefore, the uhpA gene mutants strain E. piscicida ΔuhpA was constructed to elucidate the functions of Glu6P and the uhpA gene in E. piscicida. The results demonstrated that Glu6P significantly increased the gene expression of uhpC/uhpB/uhpA than without adding Glu6P in the culture. The gene expression of uhpC and uhpB was down regulated in the mutant strain than that of in the wild type strain. E. piscicida ΔuhpA exhibited an increase in virulence compared to that of E. piscicida EIB202 [LD50 value: (3.98 × 106 CFU/fish) and LD50 value: (1.45 × 107 CFU/fish) respectively]. Besides, although TNF-α did not show significant differences (p > 0.05) in the spleen of tilapia infected with ΔuhpA and EIB202 in the whole observed period, the gene expression of IL-1β and TGF-β in the spleen of tilapia infected with ΔuhpA showed significantly higher (p < 0.05) than that of in tilapia infected with EIB202. Meanwhile, the gene expression of IL-1β and TGF-β in spleen of tilapia infected with ΔuhpA showed significantly higher (p < 0.05) than that of in fish infected with EIB202 when zebrafish used as the control in the whole observed period. All these results suggested that Glu6P up-regulated the gene expression of uhpC/uhpB/uhpA; most important, the uhpA gene deletion in E. piscicida down-regulated the gene expression of uhpC and uhpB, enhanced its pathogenicity and its role in inducing the inflammatory cytokine responses in tilapia.